Light source device and system for calcium imaging and optogenetics

Abstract

The invention relates to a light source device for calcium imaging and photoheredity. The light source device comprises a first light-emitting chip, a second light-emitting chip, a first LED lamp wick, a second LED lamp wick, a third LED lamp wick and a fourth LED lamp wick. The first light-emitting chip controls the first LED lamp wick and the third LED lamp wick to cooperatively emit first exciting light so as to excite the GCaMP calcium indicator; the second light-emitting chip controls the second LED lamp wick and the fourth LED lamp wick to cooperatively emit second exciting light so as to excite ChrimsonR photosensitive protein; the first LED lamp wick, the second LED lamp wick, the third LED lamp wick and the fourth LED lamp wick are diagonally arranged on the same plane of the substrate; wherein the first LED lamp wick and the third LED lamp wick which emit the first exciting light are distributed diagonally, and the second LED lamp wick and the fourth LED lamp wick which emit the second exciting light are distributed diagonally. The light source device disclosed by the invention can emit exciting light with two different wavelengths, so that the requirement on light source switching is reduced, and the stability of a light path is greatly improved, thereby improving the accuracy and repeatability of data.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical imaging, and in particular relates to a light source device and system for calcium imaging and optogenetics. Background Art

[0002] As neuroscience research continues to deepen, calcium imaging and optogenetics have become key means to explore the mysteries of neuronal activity. Calcium imaging reflects neuronal activity by monitoring changes in calcium ion concentration, while optogenetics uses light to precisely control neurons.

[0003] However, existing technologies have many limitations: most traditional calcium imaging and optogenetics devices rely on single-wavelength light sources, and experiments require frequent switching of equipment or the addition of additional light sources, which is not only cumbersome and costly, but also easily affects accuracy due to unstable optical paths. Moreover, conventional light sources are difficult to meet the requirements of complex experiments for light uniformity, intensity, and wavelength diversity. At the same time, the wires used in optogenetics experiments often cannot take into account the requirements of high current carrying capacity and system stability and flexibility, which limits the efficient conduct of experiments and innovative breakthroughs.

[0004] Therefore, the present invention provides a light source device and system for calcium imaging and optogenetics to solve the above technical problems. Summary of the invention

[0005] In response to the above problems, the purpose of the present invention is to provide a light source device and system for calcium imaging and optogenetics, which can emit excitation light of two different wavelengths, reduce the need for light source switching, greatly improve the stability of the optical path, and thus improve the accuracy and repeatability of the data.

[0006] The present invention provides a light source device for calcium imaging and optogenetics, which consists of a first light-emitting chip, a second light-emitting chip, a first LED lamp wick, a second LED lamp wick, a third LED lamp wick and a fourth LED lamp wick; the first light-emitting chip is connected to the first LED lamp wick and the third LED lamp wick through an extremely fine coaxial line, and is used to control the first LED lamp wick and the third LED lamp wick to cooperatively emit a first excitation light to excite a GCaMP calcium indicator; the second light-emitting chip is connected to the second LED lamp wick and the fourth LED lamp wick through an extremely fine coaxial line, and is used to control the second LED lamp wick and the fourth LED lamp wick to cooperatively emit a second excitation light to excite a ChrimsonR photosensitive protein.

[0007] Preferably, the first LED wick, the second LED wick, the third LED wick and the fourth LED wick are diagonally arranged on the same plane of the substrate; wherein the first LED wick and the third LED wick emitting the first excitation light are diagonally distributed, and the second LED wick and the fourth LED wick emitting the second excitation light are diagonally distributed.

[0008] Preferably, the first excitation light is blue light, and the wavelength range of the first excitation light is 450nm-490nm; the second excitation light is yellow-green light, and the wavelength range of the second excitation light is 590nm-650nm.

[0009] Preferably, the cross-section of the very fine coaxial line is circular; and the very fine coaxial line comprises an inner conductor, an insulating layer arranged on the outer periphery of the inner conductor, and an outer conductive structure arranged on the outer periphery of the insulating layer.

[0010] Preferably, the outer conductor structure includes an outer conductor formed by weaving a metal material and an outer sheath arranged on the outer periphery of the outer conductor.

[0011] The present invention also provides a system for calcium imaging and optogenetics, which includes an imaging device and the above-mentioned light source device, wherein the imaging device is used to receive the fluorescent signal generated by the light source device exciting the calcium indicator in the sample, and convert the fluorescent signal into an optical image signal that can be used for analysis and processing, so as to accurately capture the weak changes in the fluorescent signal, thereby realizing high-resolution detection and imaging of the changes in the calcium ion concentration in the sample.

[0012] Preferably, the imaging device comprises a housing, a lens module disposed in the housing, and a CMOS module disposed on the top of the housing; wherein the lens module comprises an excitation light filter, an achromatic lens, a beam splitter, a focusing lens, and a fluorescence filter;

[0013] The excitation light filter is arranged on the outgoing light path of the light source device, and is used to filter the first excitation light emitted by the light source device to filter out light within a specific wavelength range;

[0014] The achromatic lens is located on the outgoing light path of the excitation light filter, and is used to focus and perform chromatic aberration correction on the filtered first excitation light;

[0015] The beam splitter is used to receive the first excitation light processed by the achromatic lens, and after reflecting the first excitation light to the sample area, receives the fluorescence signal generated by the excitation of the sample area, and can allow the fluorescence signal to pass through;

[0016] The focusing lens is arranged on the optical path of the fluorescence signal transmitted by the beam splitter, and is used to focus the fluorescence signal;

[0017] The fluorescence filter is located on the outgoing light path of the focusing lens and is used to filter the fluorescence signal to obtain the fluorescence signal in a specific wavelength range;

[0018] The CMOS module is located on the outgoing light path of the fluorescent filter, and is used to receive and detect the fluorescent signal filtered by the fluorescent filter, and convert the fluorescent signal into an optical image signal that can be analyzed and processed.

[0019] Compared with the related art, the present invention provides a light source device for calcium imaging and optogenetics, comprising a first light-emitting chip, a second light-emitting chip, a first LED wick, a second LED wick, a third LED wick and a fourth LED wick; the first light-emitting chip controls the first LED wick and the third LED wick to cooperatively emit a first excitation light to excite the GCaMP calcium indicator; the second light-emitting chip controls the second LED wick and the fourth LED wick to cooperatively emit a second excitation light to excite the ChrimsonR photosensitive protein; the first LED wick, the second LED wick, the third LED wick and the fourth LED wick are arranged diagonally on the same plane of the substrate; wherein the first LED wick and the third LED wick emitting the first excitation light are diagonally distributed, and the second LED wick and the fourth LED wick emitting the second excitation light are diagonally distributed. In the above structure, the light source device can emit excitation light of two different wavelengths, which reduces the need for light source switching, greatly improves the stability of the optical path, and thus improves the accuracy and repeatability of the data. In addition, the first LED wick, the second LED wick, the third LED wick and the fourth LED wick in the light source device are arranged diagonally on the same plane of the substrate, and the wicks emitting excitation light of the same wavelength are distributed diagonally, which can help achieve more uniform color mixing, thereby reducing color gradients or shadows, and can also bring better heat distribution, so that heat is more evenly distributed on the substrate, which helps to extend the service life of the LED wicks and improve reliability. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a structural block diagram of a light source device for calcium imaging and optogenetics of the present invention;

[0021] Figure 2 It is a schematic diagram of the three-dimensional structure of the first LED lamp core, the second LED lamp core, the third LED lamp core and the fourth LED lamp core in the present invention;

[0022] Figure 3 It is a schematic diagram of the three-dimensional structure of the ultra-thin coaxial line in the present invention;

[0023] Figure 4A schematic diagram of the three-dimensional structure of a system for calcium imaging and optogenetics of the present invention;

[0024] Figure 5 This is a schematic diagram of optical path imaging of a system for calcium imaging and optogenetics of the present invention. DETAILED DESCRIPTION

[0025] The present invention provides a light source device and system for calcium imaging and optogenetics, aiming to solve the problem that most existing calcium imaging and optogenetics equipment rely on a single wavelength light source, and the equipment needs to be frequently switched or additional light sources need to be added during the experiment, which easily leads to the problem that the accuracy is affected by the unstable light path.

[0026] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Please refer to the attached Figure 1-5 As shown, the present invention provides a light source device 100 for calcium imaging and optogenetics, comprising a first light-emitting chip 10, a second light-emitting chip 20, a first LED lamp core 1, a second LED lamp core 2, a third LED lamp core 3 and a fourth LED lamp core 4; the first light-emitting chip 10 is connected to the first LED lamp core 1 and the third LED lamp core 3 through an extremely fine coaxial line 30, and is used to control the first LED lamp core 1 and the third LED lamp core 3 to cooperatively emit a first excitation light to excite a GCaMP calcium indicator; the second light-emitting chip 20 is connected to the second LED lamp core 1 through an extremely fine coaxial line 30. The wick 2 is connected to the fourth LED wick 4, and is used to control the second LED wick 2 and the fourth LED wick 4 to collaboratively emit the second excitation light to excite the ChrimsonR photosensitive protein; the first LED wick 1, the second LED wick 2, the third LED wick 3 and the fourth LED wick 4 are diagonally arranged on the same plane of the substrate; wherein the first LED wick 1 and the third LED wick 3 emitting the first excitation light are diagonally distributed, and the second LED wick 2 and the fourth LED wick 4 emitting the second excitation light are diagonally distributed.

[0028] In this embodiment, the first excitation light is blue light, and the wavelength range of the first excitation light is 450nm-490nm; the second excitation light is yellow-green light, and the wavelength range of the second excitation light is 590nm-650nm. Of course, it is not limited to this, and the colors of the first excitation light and the second excitation light can be adjusted according to experimental requirements.

[0029] In the above structure, the first LED wick 1, the second LED wick 2, the third LED wick 3 and the fourth LED wick 4 in the light source device are diagonally arranged on the same plane of the substrate, and the LED wicks emitting excitation light of the same wavelength are diagonally distributed, which has the following advantages: 1. It improves the light output efficiency, reduces light loss, and provides a stronger excitation light source. 2. It can help achieve more uniform color mixing, thereby reducing color gradients or shadows; and it can achieve more uniform lighting, which not only makes the fluorescent probe respond more consistently to different wavelengths and reduces signal fluctuations, but also eliminates the chromatic aberration problem that may be caused by differences in color distribution, and improves the color consistency of the image; 3. It improves the resolution of imaging and supports more detailed observation of intracellular structures; 4. There are slight differences in thermal distribution between different types of LED wicks. The diagonal layout brings better thermal distribution, so that heat is more evenly distributed on the substrate, which helps to extend the service life of the LED wick and improve reliability. From certain angles, the diagonal arrangement can reduce the phenomenon of a certain color dominance. In visual perception, this diagonal arrangement reduces the impact of wicks of different colors on the viewing angle, improves the display effect, and makes the color performance more consistent when viewed from most angles.

[0030] It should be noted that the two colors of excitation light in the light source device are controlled and emitted by the first light-emitting chip 10 and the second light-emitting chip 20, respectively, so that the two LED wicks that emit the two colors of excitation light can be independently controlled, which is highly flexible and can adjust the light source according to experimental requirements to adapt to different fluorescent probes and experimental conditions, and provide higher light source uniformity, which is particularly critical in calcium imaging, because the intensity and quality of the fluorescent signal directly depend on the uniformity of the excitation light; and the wavelength and intensity of the excitation light emitted by the LED wick can be accurately controlled by the first light-emitting chip 10 and the second light-emitting chip 20, so that researchers can detect the dynamic changes of calcium ions in cells. Therefore, the uniform illumination of the LED wick can not only improve the accuracy of the data, but also prevent signal deviation, ensuring that the obtained imaging data has high resolution and high reliability.

[0031] In this embodiment, the cross section of the ultra-fine coaxial line 30 is circular; and the ultra-fine coaxial line 30 includes an inner conductor 301, an insulating layer 302 arranged on the outer periphery of the inner conductor 301, and an outer conductive structure 303 arranged on the outer periphery of the insulating layer 302; and the outer conductive structure 303 includes an outer conductor 3031 woven from a metal material and an outer sheath 3032 arranged on the outer periphery of the outer conductor. The setting of this structure effectively reduces the resistance when the current passes through, improves the transmission efficiency, reduces the energy loss, and can withstand high temperatures, effectively prevents the safety hazards caused by overheating, provides reliable current transmission, enables researchers to obtain more stable light output in optogenetic experiments, ensures that the light-sensitive channel protein is excited to achieve the ideal light intensity, thereby improving the success rate of the experiment, and also broadens the application scope of optogenetic technology, providing a solid technical foundation for precise biological regulation. In addition, the ultra-fine coaxial line 30 has better flexibility, making it more flexible to arrange in the experimental environment and able to adapt to complex experimental needs.

[0032] It is worth mentioning that the present invention provides a system for calcium imaging and optogenetics, including an imaging device 200 and the above-mentioned light source device 100, wherein the imaging device 200 is used to receive the fluorescence signal generated by the light source device 100 exciting the calcium indicator in the sample, and convert the fluorescence signal into an optical image signal for analysis and processing, so as to accurately capture the weak fluorescence signal changes, thereby realizing high-resolution detection and imaging of the changes in calcium ion concentration in the sample.

[0033] In this embodiment, the imaging device 200 includes a shell 201, a lens module 202 arranged in the shell 201, and a CMOS module 203 arranged on the top of the shell 201; wherein the lens module 202 includes an excitation light filter 2021, an achromatic lens 2022, a spectrometer 2023, a focusing lens 2024 and a fluorescence filter 2025.

[0034] The excitation light filter 2021 is disposed on the outgoing light path of the light source device 100, and is used to filter the first excitation light emitted by the light source device 100 to filter out light within a specific wavelength range;

[0035] The achromatic lens 2022 is located on the outgoing light path of the excitation light filter 2021, and is used to focus and perform chromatic aberration correction on the filtered first excitation light;

[0036] The beam splitter 2023 is used to receive the first excitation light processed by the achromatic lens 2022, and after reflecting the first excitation light to the sample area, receive the fluorescence signal generated by the excitation of the sample area, and can allow the fluorescence signal to pass through;

[0037] The focusing lens 2024 is disposed on the optical path of the fluorescence signal transmitted by the beam splitter 2023, and is used to focus the fluorescence signal;

[0038] The fluorescence filter 2025 is located on the outgoing light path of the focusing lens 2024 and is used to filter the fluorescence signal to obtain the fluorescence signal in a specific wavelength range;

[0039] The CMOS module 203 is located on the outgoing light path of the fluorescent filter 2025, and is used to receive and detect the fluorescent signal filtered by the fluorescent filter 2025, and convert the fluorescent signal into an optical image signal that can be analyzed and processed.

[0040] Compared with the related art, the present invention provides a light source device for calcium imaging and optogenetics, comprising a first light-emitting chip, a second light-emitting chip, a first LED wick, a second LED wick, a third LED wick and a fourth LED wick; the first light-emitting chip controls the first LED wick and the third LED wick to cooperatively emit a first excitation light to excite the GCaMP calcium indicator; the second light-emitting chip controls the second LED wick and the fourth LED wick to cooperatively emit a second excitation light to excite the ChrimsonR photosensitive protein; the first LED wick, the second LED wick, the third LED wick and the fourth LED wick are arranged diagonally on the same plane of the substrate; wherein the first LED wick and the third LED wick emitting the first excitation light are diagonally distributed, and the second LED wick and the fourth LED wick emitting the second excitation light are diagonally distributed. In the above structure, the light source device can emit excitation light of two different wavelengths, which reduces the need for light source switching, greatly improves the stability of the optical path, and thus improves the accuracy and repeatability of the data. In addition, the first LED wick, the second LED wick, the third LED wick and the fourth LED wick in the light source device are arranged diagonally on the same plane of the substrate, and the wicks emitting excitation light of the same wavelength are distributed diagonally, which can help achieve more uniform color mixing, thereby reducing color gradients or shadows, and can also bring better heat distribution, so that heat is more evenly distributed on the substrate, which helps to extend the service life of the LED wicks and improve reliability.

[0041] The above-described embodiments should be understood as illustrative rather than limiting the scope of protection of the present invention, which is subject to the claims. For those skilled in the art, some non-essential improvements and adjustments made to the present invention still fall within the scope of protection of the present invention without departing from the essence and scope of the present invention.

Claims

1. A light source device for calcium imaging and optogenetics, characterized in that: The light source device is composed of a first light-emitting chip, a second light-emitting chip, a first LED lamp core, a second LED lamp core, a third LED lamp core and a fourth LED lamp core; the first light-emitting chip is connected to the first LED lamp core and the third LED lamp core through an extremely fine coaxial line, and is used to control the first LED lamp core and the third LED lamp core to cooperatively emit a first excitation light to excite the GCaMP calcium indicator; The second light-emitting chip is connected to the second LED lamp core and the fourth LED lamp core through an extremely fine coaxial line, and is used to control the second LED lamp core and the fourth LED lamp core to cooperatively emit a second excitation light to excite the ChrimsonR photosensitive protein.

2. A light source device for calcium imaging and optogenetics according to claim 1, characterized in that: The first LED lamp core, the second LED lamp core, the third LED lamp core and the fourth LED lamp core are arranged diagonally on the same plane of the substrate; wherein the first LED lamp core and the third LED lamp core emitting the first excitation light are diagonally distributed, and the second LED lamp core and the fourth LED lamp core emitting the second excitation light are diagonally distributed.

3. A light source device for calcium imaging and optogenetics according to claim 1, characterized in that: The first excitation light is blue light, and the wavelength range of the first excitation light is 450nm-490nm; the second excitation light is yellow-green light, and the wavelength range of the second excitation light is 590nm-650nm.

4. A light source device for calcium imaging and optogenetics according to claim 1, characterized in that: The cross section of the very fine coaxial line is circular; and the very fine coaxial line comprises an inner conductor, an insulating layer arranged on the outer periphery of the inner conductor, and an outer conductive structure arranged on the outer periphery of the insulating layer.

5. A light source device and system for calcium imaging and optogenetics according to claim 4, characterized in that: The outer conductor structure comprises an outer conductor formed by weaving a metal material and an outer sheath arranged at the outer periphery of the outer conductor.

6. A system for calcium imaging and optogenetics, characterized in that The system comprises an imaging device and a light source device as described in any one of claims 1 to 5 above, wherein the imaging device is used to receive a fluorescent signal generated by the light source device exciting a calcium indicator in a sample, and convert the fluorescent signal into an optical image signal that can be analyzed and processed, so as to accurately capture weak changes in the fluorescent signal, thereby achieving high-resolution detection and imaging of changes in calcium ion concentration in the sample.

7. A system for calcium imaging and optogenetics according to claim 6, characterized in that: The imaging device comprises a housing, a lens module arranged in the housing, and a CMOS module arranged on the top of the housing; wherein the lens module comprises an excitation light filter, an achromatic lens, a beam splitter, a focusing lens, and a fluorescence filter; The excitation light filter is arranged on the outgoing light path of the light source device, and is used to filter the first excitation light emitted by the light source device to filter out light within a specific wavelength range; The achromatic lens is located on the outgoing light path of the excitation light filter, and is used to focus and perform chromatic aberration correction on the filtered first excitation light; The beam splitter is used to receive the first excitation light processed by the achromatic lens, and after reflecting the first excitation light to the sample area, receives the fluorescence signal generated by the excitation of the sample area, and can allow the fluorescence signal to pass through; The focusing lens is arranged on the optical path of the fluorescence signal transmitted by the beam splitter, and is used to focus the fluorescence signal; The fluorescence filter is located on the outgoing light path of the focusing lens and is used to filter the fluorescence signal to obtain the fluorescence signal in a specific wavelength range; The CMOS module is located on the outgoing light path of the fluorescent filter, and is used to receive and detect the fluorescent signal filtered by the fluorescent filter, and convert the fluorescent signal into an optical image signal that can be analyzed and processed.

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